Abstract

Pomegranate is a widely distributed species in many tropical and subtropical regions. Pomegranate is consumed as fresh fruit or raw material to produce various products such as juice, syrup, jams, and wine. DNA extraction is a critical way to study the genetic diversity and phylogeny of the pomegranate species. Hence, chelex is an accessible and safe method to extract the DNA of pomegranate. So far, chelex was not used for DNA extraction in this plant and therefore it has been evaluated for DNA isolation in South African pomegranate for the firs time. This study was carried out at Genetic lab at the University of Limpopo during 2021to compare the two different extraction DNA methods from pomegranate (P. granatum). The DNA from the pomegranate was extracted using Chelex 100 “method 1” (whole night incubation at 56°C) and “method 2” (Ten minutes incubation at 95°C) using the fresh leaves of the pomegranate. For this purpose, four samples of P. granatum from northen parts of South Africa were studied. Quantitative and qualitative parameters were assessed using a spectrophotometer. The PCR reaction with primers for 28S were used on all samples to confirm and evaluate the extracted DNA. The results for the spectrophotometer indicated that the highest quality of extracted DNA was in “method 2” (1.45–1.69). However, the protein (1.41–1.52 mg/mL) was detected through “method 1” in the tested samples. The qualitative and quantitative tests for PCR processing realised that the total DNA extracted using “method 2” had improved quality than “method 1”. Amplification of samples using 28S rDNA primer showed a higher concentration and purity of DNA extracted using “method 2”. In conclusion, both methods yield enough DNA, however, “method 2” yielded higher quality DNA in a less time.